Identification of potential transcription factors that enhance human iPSC generation

Swaidan, Nuha T.; Salloum-Asfar, Salam; Palangi, Freshteh; Errafii, Khaoula; Soliman, Nada; Aboughalia, Ahmed T.M.; Wali, Abdul Haseeb S.; Abdulla, Sara; Emara, Mohamed M.

doi:10.1038/s41598-020-78932-9

Cited by 13 publications

(15 citation statements)

References 49 publications

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“…1j). Engineered UVC also showed similar expression to control iPS cells for genes (DNC, Vimentin, HES5 and GATA6) that are known to increase in expression as iPS cells differentiate into mesoderm, endoderm and ectoderm liniage, confirming the UVC have a consistent undifferentiated, iPS cell gene expression profile, morphology, and growth characteristics 47,48 Taken together, these data demonstrate that the CRISPR engineered UVC has the capacity to deliver abundant, full-length spike protein antigen in the context of an irradiated, apoptotic cellular vehicle.…”

Section: Rapid Growth Kinetics Of Engineered Uvcmentioning

confidence: 63%

A Novel CRISPR-Engineered, Stem Cell-Derived Cellular Vaccine

Chakraborty

Chandrashekar

Sidaway

et al. 2021

Preprint

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COVID-19 has forced rapid clinical translation of novel vaccine technologies, principally mRNA vaccines, that have resulted in meaningful efficacy and adequate safety in response to the global pandemic. Notwithstanding this success, there remains an opportunity for innovation in vaccine technology to address current limitations and meet the challenges of inevitable future pandemics. We describe a universal vaccine cell (UVC) rationally designed to mimic the natural physiologic immunity induced post viral infection of host cells. Induced pluripotent stem cells were CRISPR engineered to delete MHC-I expression and simultaneously overexpress a NK Ligand adjuvant to increase rapid cellular apoptosis which was hypothesized to enhance viral antigen presentation in the resulting immune microenvironment leading to a protective immune response. Cells were further engineered to express the parental variant WA1/2020 SARS-CoV-2 spike protein as a representative viral antigen prior to irradiation and cryopreservation. The cellular vaccine was then used to immunize non-human primates in a standard 2-dose, IM injected prime + boost vaccination with 1e8 cells per 1 ml dose resulting in robust neutralizing antibody responses (1e3 nAb titers) with decreasing levels at 6 months duration. Similar titers generated in this established NHP model have translated into protective human neutralizing antibody levels in SARS-Cov-2 vaccinated individuals. Animals vaccinated with WA1/2020 spike antigens were subsequently challenged with 1.0 x 105 TCID50 infectious Delta (B.1.617.2) SARS-CoV-2 in a heterologous challenge which resulted in an approximately 3-log order decrease in viral RNA load in the lungs. These heterologous viral challenge results reflect the ongoing real-world experience of original variant WA1/2020 spike antigen vaccinated populations exposed to rapidly emerging variants like Delta and now Omicron. This cellular vaccine is designed to be a rapidly scalable cell line with a modular poly-antigenic payload to allow for practical, large-scale clinical manufacturing and use in an evolving viral variant environment. Human clinical translation of the UVC is being actively explored for this and potential future pandemics.

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Section: Rapid Growth Kinetics Of Engineered Uvcmentioning

confidence: 63%

A Novel CRISPR-Engineered, Stem Cell-Derived Cellular Vaccine

Chakraborty

Chandrashekar

Sidaway

et al. 2021

Preprint

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“…Engineered UVCs also showed similar expression to control iPSCs for genes (DCN, vimentin, HES5, and GATA6) that are known to increase in expression as iPSCs differentiate into mesoderm, endoderm, and ectoderm lineages, confirming that the UVCs have a consistent undifferentiated iPSC gene expression profile, morphology, and growth characteristics. 47 , 48 …”

Section: Resultsmentioning

confidence: 99%

A genetically engineered, stem-cell-derived cellular vaccine

Cooper¹,

Sidaway²,

Chandrashekar

et al. 2022

Cell Reports Medicine

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“…Nanog and Myc, along with Krüppel-like factor 4, are naïve pluripotent markers that drive the transition of EpSCs to ESCs ( 39 ). Moreover, Nanog and Myc gene expression are independent of each other in the transcriptional regulatory pathways and do not directly regulate each other, but the Nanog and Myc both stimulate cell proliferation ( 28 ).…”

Section: Discussionmentioning

confidence: 99%

Epidermal stem cells participate in the repair of scalds via Nanog and Myc regulation

et al. 2022

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epidermal stem cells (epScs) with high expression of regulatory factor nanog can promote wound healing. The aim of the present study was to investigate the effectiveness and mechanism of epidermal stem cells (epScs) in healing scalds and the underlying molecular mechanism. Mouse epScs were isolated from skin tissues and cultured in vitro. First, the proliferative ability of epScs was determined via the upregulation and downregulation of nanog expression levels in epScs using the MTS-assay. Second, a wound healing assay of the epScs with different nanog expression levels was performed to investigate cell migratory capacities. Third, the protein expression levels of various proteins in epScs with nanog overexpression or knockdown, were determined. Finally, the transfected epScs were applied to the rat scald model to observe their effect on scald healing. Subsequently, wound scores, re-epithelialization and capillary density were determined histologically. The results demonstrated that nanog overexpression enhanced the proliferative ability of epScs via cellular (c)-Myc. Moreover, the lV-nanog group of epScs with increased nanog expression levels exhibited improved healing abilities in the wound healing test than control group. using western blotting, it was demonstrated that epScs that were transfected with a nanog-overexpression vector expressed high nanog protein expression levels, whereas small interfering rna-nanog-transfected epScs exhibited low nanog protein expression levels. Furthermore, c-Myc expression was synchronized with nanog expression. it was also revealed that as the expression levels of c-Myc increased, p53 expression levels also increased. in the rat scald model, nanog-overexpressing epScs enhanced wound closure and re-epithelialization. The epScs with nanog knockdown exhibited the opposite effects. The present study therefore indicated that nanog may have a positive effect on scald healing in rats, which supports its use in epSc-based treatments against scalds. Furthermore, it was suggested that c-Myc potentially serves a key role in this process and that this process avoids cancerization by relying on the supervision of p53.

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Identification of potential transcription factors that enhance human iPSC generation

Cited by 13 publications

References 49 publications

A Novel CRISPR-Engineered, Stem Cell-Derived Cellular Vaccine

A Novel CRISPR-Engineered, Stem Cell-Derived Cellular Vaccine

A genetically engineered, stem-cell-derived cellular vaccine

Epidermal stem cells participate in the repair of scalds via Nanog and Myc regulation

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